The principal long-term objectives of this proposal are to develop an efficient novel assay methodology for the detection of the category B biothreat agents, Burkholderia mallei (the causative agent of glanders) and Burkholderia pseudomallei (the causative agent of melioidosis). Infections are particularly debilitating and can lead to abscesses in the lungs and other organs with accompanying septicemia. These organisms are particularly dangerous as there are no licensed vaccines and treatment with antibiotics is prolonged and often ineffective. Improved, sensitive methods for whole organism detection will be of great value in protecting both military and civilian populations, particularly as the infection would most likely be in the form of an aerosol. Furthermore, development of detection methods to identify Burkholderia relative to other Gram-negative organisms and to distinguish between species would be of significant value in ensuring that correct countermeasures could be rapidly implemented. We propose to produce surface-associated molecules and membranes from Burkholderia organisms, to develop high-affinity aptamers and antibodies that can bind to these targets, and to adapt Luminex dye- based technology to generate spectrally sensitive assays for the detection of Burkholderia species. Additionally, we will assess the therapeutic potential of reagents generated as a consequence of these studies The specific aims are: 1. To produce candidate diagnostic development targets from pathogenic Burkholderia species. 2. To generate high affinity aptamers and antibodies against these targets. 3. To develop robust assays for the detection of these targets and optiise the assays also for detection of whole organisms. 4. To assess the potential therapeutic value of reagents generated within this project. The principal objective of this work is to develop new diagnostic assays for the rapid detection of the microbial pathogens, Burkholderia mallei and Burkholderia pseudomallei. The development of a rapid, more robust method for detecting these organisms will be of value for both the military (as the organisms are biothreat agents) and in clinics in the developing world (e.g., Southeast Asia).